甘肃天水产花椒果实资源化学研究

发布时间：2018-08-29 09:34

【摘要】：芸香科花椒属植物花椒Zanthoxylum bungeanum Maxim.为药食两用植物,其干燥成熟果皮入药称花椒(Zanthoxyli Pericarpium),具有温中止痛、杀虫止痒的功效,为中医临床常用温里药味,常用于脘腹冷痛、虫积腹痛、呕吐泄泻、蛔虫病等症的治疗;亦可外用治疗湿疹瘙痒。花椒种子入药称椒目(Zanthoxyli Semen),具有渗水利湿、祛痰平喘之功,临床主要用于水肿胀满、哮喘等症的治疗。我国花椒资源丰富,产量巨大。现代研究显示,花椒果实中含有多种类型的资源性化学成分,在医药、食品、保健品和化妆品等多个领域应用广泛。在对相关文献进行整理分析,系统综述我国花椒属药用植物资源研究现状的基础上,本研究选择甘肃天水地区产花椒成熟果实为研究对象,基于中药资源化学的研究思路与方法,立足于资源多途径、多层次利用原则,以花椒资源的综合利用和产业化发展为目的,对花椒果实开展了资源性化学成分分析与评价,并就其潜在资源价值与产品开发作了初步探索,以期为花椒果实资源的综合利用与产业化开发提供科学依据,也为提升花椒资源的利用效率和效益提供支撑。1.花椒果实资源性化学成分分析与评价为明确花椒果实中资源性物质的组成及分布特征,充分释放花椒果实资源价值,本研究分别采用气相色谱-质谱联用法(GC-MS)、高效液相色谱法(HPLC-PAD/ELSD)、超高效液相色谱-质谱联用法(UPLC-TQ/MS)等技术手段,对花椒成熟果实的果皮、种子及种子油中所含的挥发油类、酚酸类、黄酮类等与药用功效相关的资源性活性物质,以及脂肪酸类、糖类、蛋白质及氨基酸类、核苷及碱基类等与营养保健相关的资源性物质进行了系统分析与评价。(1)挥发油类资源性化学成分分析结果显示,花椒果皮、种子中存在多种不同类型的挥发性成分,其中萜类及其含氧衍生物为挥发油中的主要组分,乙酸芳樟酯在花椒果实各部位挥发油所占比例相对较高。比较所检测花椒果实不同部位,发现果皮中的挥发油类成分较种子组成相对更为丰富,主要成分的含量也相对较高。(2)黄酮类和酚酸类资源性化学成分分析结果表明,花椒果实中的黄酮类和酚酸类资源性化学成分主要分布于果皮部位,其主要组成成分绿原酸、槲皮素-3-O-芸香糖苷、金丝桃苷、槲皮素-3-O-葡萄糖苷和槲皮苷的含量分别为2.15、1.99、3.94、1.69、2.04 mg/g;而花椒种子和花椒种子油中未检测到上述酚酸类和黄酮类资源性化学成分。(3)脂肪酸类资源性化学成分研究显示,花椒果皮和种子中的脂肪酸总量分别为108.42mg/g、331.63mg/g;由花椒种子压榨制备的种子油中脂肪酸总量可达966.04mg/g,单不饱和及多不饱和脂肪酸总量约占其总量的78.95%,其中尤以油酸、亚油酸和亚麻酸含量相对较高,其质量分数分别达293.35mg/g、246.56mg/g、184.41mg/g。(4)糖类资源性化学成分分析结果显示,花椒果皮与种子中可溶性多糖总含量分别为96.33mg/g、19.18mg/g;单糖/寡糖总含量分别为9.71mg/g和3.78mg/g。可见,花椒果实中的糖类组分主要分布于果皮,而中性多糖为花椒果实主要多糖类型。(5)核苷及碱基类资源性化学成分分析结果显示,花椒果皮与种子中的核苷及碱基类化学成分在组成与含量上均存在差异,花椒果皮与种子中核苷及碱基类成分的总含量分别为2.14mg/g、3.94mg/g,果皮与种子中均以尿苷含量最高分别为0.47mg/g、1.22mg/g,腺苷、肌苷和胸腺嘧啶在花椒果皮和种子中所占比例也相对较高。(6)总蛋白及氨基酸类资源性化学成分分析结果显示,花椒果皮中总蛋白的含量(92.32mg/g)远高于种子(10.97mg/g);果皮中水解氨基酸总量约为52.90mg/g,其中以脯氨酸的含量为最高(9.95mg/g);种子中水解氨基酸总量约为4.66mg/g,以谷氨酸所占比例为最高(1.03mg/g)。花椒果实中游离氨基酸测定结果表明,其在花椒果皮中的总量约为11.02mg/g,在花椒种子中含量相对较低,仅为0.57mg/g,且果皮和种子中游离氨基酸均为精氨酸和脯氨酸含量相对较高。2.花椒果实资源化利用研究(1)采用卵清白蛋白复合寒冷与过劳刺激致寒饮蕴肺哮喘大鼠模型,对花椒种子水提物及种子油的效应特征进行评价研究,结果显示,花椒种子水提物与种子油对模型大鼠血液炎性细胞、炎症因子、肺组织病理学损伤均有一定程度的干预作用,各剂量花椒种子水提物给药组及种子油给药组哮喘大鼠均有趋于正常化的趋势,表明花椒种子具有一定的防治哮喘作用。(2)基于代谢组学技术对花椒种子干预寒饮蕴肺哮喘大鼠的作用机制研究发现,模型组大鼠与正常组大鼠比较,在其血清和尿液中共鉴定28个潜在生物标志物。其中,血清中鉴定的潜在生物标志物涉及的代谢通路中,亚油酸代谢、花生四烯酸代谢、乙醛酸和二羧酸代谢和甘油磷脂代谢通路影响值相对较高;大鼠尿液中所鉴定的潜在生物标志物涉及的代谢通路中,谷氨酰胺与谷氨酸代谢,戊糖与葡萄糖醛酸生物转化,丙氨酸、天门冬氨酸和谷氨酸代谢,烟酸盐与烟酰胺代谢,鞘脂类代谢、精氨酸与脯氨酸代谢通路影响值相对较高。对花椒种子水提物和种子油药物治疗组大鼠的血清与尿液中的上述潜在标志性代谢物分析发现,试药通过影响亚油酸代谢、甘油磷脂代谢、戊糖和葡萄糖醛酸酯生物转化、烟酸盐和烟酰胺代谢这几条代谢途径对哮喘进行干预治疗。(3)对花椒种子油、花椒果皮挥发油与酰胺部位体外抗氧化活性研究结果表明,酰胺部位对DPPH和ABTS表现出较强的清除能力,其IC50(半数抑制浓度值)分别为1.55mg/m L、0.23 mg/m L;挥发油对ABTS自由基的清除效果显著,IC50值为1.07 mg/m L,挥发油和种子油对DPPH自由基的清除能力较酰胺部位弱,IC50值分别为19.36 mg/m L和16.38 mg/m L;花椒挥发油在FRAP试验中表现还原能力最强,其次为酰胺部位,三组样品的抗氧化水平在FeSO4线性范围内呈现出一定的量效相关性。
[Abstract]:Zanthoxylum bungeanum Maxim, a Rutaceae Zanthoxylum plant, is an edible and medicinal plant. Its dried and ripe peel is called Zanthoxyli Pericarpium. It has the effect of stopping pain and killing itching. It is commonly used in traditional Chinese medicine clinic. It is used in the treatment of cold abdominal pain, epigastric pain, vomiting and diarrhea, ascariasis, etc. Zanthoxyli Semen is used to treat eczema and itching. Zanthoxyli Semen has the functions of water permeation, dampness elimination, expectoration and relieving asthma. It is mainly used in the treatment of edema, asthma and other symptoms. China is rich in Zanthoxyli resources and has a huge production. Modern research shows that Zanthoxyli Semen contains a variety of resource chemical components in the fruit, in medicine, food, health care. It is widely used in many fields, such as cosmetics and so on. On the basis of sorting out and analyzing the related literature and systematically reviewing the research status of Zanthoxylum medicinal plant resources in China, the ripe fruit of Zanthoxylum bungeanum in Tianshui area of Gansu province was selected as the research object. Based on the research ideas and methods of Chinese medicine resources chemistry, the resources of Zanthoxylum bungeanum bungeanum bungean Based on the principle of hierarchical utilization, the chemical composition of Zanthoxylum bungeanum was analyzed and evaluated for the purpose of comprehensive utilization and industrialization development of Zanthoxylum bungeanum resources, and the potential resource value and product development were preliminarily explored in order to provide scientific basis for the comprehensive utilization and industrialization development of Zanthoxylum bungeanum resources as well as to raise the capital of Zanthoxylum bungeanum. The analysis and evaluation of the chemical constituents of Zanthoxylum bungeanum fruits were carried out to clarify the composition and distribution characteristics of the resource substances in Zanthoxylum bungeanum fruits and fully release the resource value of Zanthoxylum bungeanum fruits. Phase chromatography-mass spectrometry (UPLC-TQ/MS) and other technical means, pepper mature fruit peel, seeds and seed oil contained in the volatile oils, phenolic acids, flavonoids and other medicinal efficacy-related resource active substances, as well as fatty acids, carbohydrates, proteins and amino acids, nucleosides and alkaloids related to nutrition and health resources The results showed that there were many different types of volatile components in the peel and seeds of Zanthoxylum bungeanum. Terpenoids and their oxygen-containing derivatives were the main components of volatile oils. Linalyl acetate accounted for a relatively high proportion of volatile oils in Zanthoxylum bungeanum fruit. Comparing the different parts of Zanthoxylum bungeanum fruits, it was found that the volatile oils in the peel were more abundant than those in the seed, and the contents of the main components were also relatively high. (2) The analysis of flavonoids and phenolic acids showed that the flavonoids and phenolic acids in Zanthoxylum bungeanum fruits were mainly distributed in the peel. The contents of chlorogenic acid, quercetin-3-O-rutin, hyperoside, quercetin-3-O-glucoside and quercetin were 2.15, 1.99, 3.94, 1.69, 2.04 mg/g, respectively, while the phenolic acids and flavonoids were not detected in Zanthoxylum bungeanum seeds and Zanthoxylum bungeanum seed oils. (3) The study of fatty acid resource chemical constituents was obvious. The results showed that the total fatty acids in peel and seed of Zanthoxylum bungeanum were 108.42 mg/g and 331.63 mg/g, respectively; the total fatty acids in seed oil prepared by pressing Zanthoxylum bungeanum bungeanum seeds were 966.04 mg/g, and the total monounsaturated and polyunsaturated fatty acids accounted for 78.95% of the total fatty acids, especially oleic acid, linoleic acid and linolenic acid, with relatively high contents and their mass fractions respectively. The total contents of soluble polysaccharides in peel and seed of Zanthoxylum bungeanum were 96.33 mg / g and 19.18 mg / g respectively, and the total contents of monosaccharide / oligosaccharides were 9.71 mg / g and 3.78 mg / g respectively. (5) Nucleosides and alkaloids in the peel and seed of Zanthoxylum bungeanum were different in composition and content. The total contents of nucleosides and alkaloids in the peel and seed of Zanthoxylum bungeanum were 2.14mg/g and 3.94mg/g, respectively. Uridine was found in the peel and seed of Zanthoxylum bungeanum bungeanum. The contents of adenosine, inosine and thymidine in peel and seed of Zanthoxylum bungeanum were 0.47 mg/g, 1.22 mg/g, respectively. (6) The analysis of total protein and amino acid resources showed that the total protein content in peel of Zanthoxylum bungeanum (92.32 mg/g) was much higher than that in seed (10.97 mg/g), and the total hydrolyzed amino acid content in peel was about 52.90 M. The content of proline was the highest (9.95 mg/g), the total amount of hydrolyzed amino acids in seeds was about 4.66 mg/g, and the proportion of glutamic acid was the highest (1.03 mg/g). The total content of free amino acids in Pepper pericarp was about 11.02 mg/g, and the content of free amino acids in pepper seed was relatively low (0.57 mg/g). The content of free amino acids in seeds was relatively high. 2. Utilization of Zanthoxylum bungeanum L. fruits was studied. (1) The effects of water extract and seed oil from Zanthoxylum bungeanum L. seeds on asthmatic rats induced by cold and overwork combined with ovalbumin were evaluated. Seed oil could interfere with inflammatory cells, inflammatory factors and lung histopathological injury in model rats to a certain extent, and the asthmatic rats tended to normalize in the water extract treatment group and the seed oil treatment group of Zanthoxylum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bungeanum bun The mechanism of Zanthoxylum bungeanum seed intervention on asthmatic rats with cold-retention lung syndrome was studied. 28 potential biomarkers were identified in serum and urine of model rats compared with normal rats. Among them, linoleic acid metabolism, arachidonic acid metabolism, glyoxylic acid and dicarboxylic acid metabolism were involved in the potential biomarkers identified in serum. Among the metabolic pathways identified in rat urine, glutamine and glutamate metabolism, pentose and glucuronic acid biotransformation, alanine, aspartate and glutamate metabolism, nicotinate and nicotinamide metabolism, sphingolipid metabolism, arginine and glutamate metabolism were involved. Proline metabolic pathways were relatively high. The analysis of the above potential markers in serum and urine of rats treated with water extract and seed oil from Zanthoxylum bungeanum seeds showed that the tested drugs affected linoleic acid metabolism, glycerol phospholipid metabolism, pentose and glucuronide ester biotransformation, nicotinate and nicotinamide metabolism. (3) In vitro antioxidant activity of Zanthoxylum bungeanum seed oil, Zanthoxylum bungeanum peel volatile oil and amide fraction showed strong scavenging ability to DPPH and ABTS, and their IC50 (half inhibitory concentration) were 1.55 mg/ml, 0.23 mg/ml, respectively. The IC50 value was 1.07 mg/ml, the scavenging ability of volatile oil and seed oil to DPPH free radical was weaker than that of amide part, IC50 value was 19.36 mg/ml and 16.38 mg/ml, respectively; the volatile oil of Zanthoxylum bungeanum showed the strongest reducing ability in FRAP test, followed by amide part, and the antioxidant level of the three groups of samples showed a certain dose-effect phase in the linear range of FeSO4. Customs.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

【参考文献】